Expression Of Cell Factors Research Articles

Pulmonary decellularized extracellular matrix (dECM) modified polyethylene terephthalate three-dimensional cell carriers regulate the proliferation and paracrine activity of mesenchymal stem cells.

Introduction: Mesenchymal stem cells (MSCs) possess a high degree of self-renewal capacity and in vitro multi-lineage differentiation potential. Decellularized materials have garnered considerable attention due to their elevated biocompatibility, reduced immunogenicity, excellent biodegradability, and the ability to partially mimic the in vivo microenvironment conducive to cell growth. To address the issue of mesenchymal stem cells losing their stem cell characteristics during two-dimensional (2D) cultivation, this study established three-dimensional cell carriers modified with lung decellularized extracellular matrix and assessed its impact on the life activities of mesenchymal stem cells. Methods: This study employed PET as a substrate material, grafting with polydopamine (PDA), and constructing a decellularized extracellular matrix (dECM) coating on its surface, thus creating the PET/PDA/dECM three-dimensional (3D) composite carrier. Subsequently, material characterization of the cellular carriers was conducted, followed by co-culturing with human umbilical cord mesenchymal stem cells in vitro, aiming to investigate the material's impact on the proliferation and paracrine activity of mesenchymal stem cells. Results and Discussion: Material characterization demonstrated successful grafting of PDA and dECM materials, and it had complete hydrophilicity, high porosity, and excellent mechanical properties. The material was rich in various ECM proteins (collagen I, collagen IV , laminin, fibronectin, elastin), indicating good biocompatibility. In long-term in vitro cultivation (14days) experiments, the PET/PDA/dECM three-dimensional composite carrier significantly enhanced adhesion and proliferation of human umbilical cord-derived mesenchymal stem cells (HUCMSCs), with a proliferation rate 1.9 times higher than that of cells cultured on tissue culture polystyrene (TCPS) at day 14. Furthermore, it effectively maintained the stem cell characteristics, expressing specific antigens for HUCMSCs. Through qPCR, Western blot, and ELISA experiments, the composite carrier markedly promoted the expression and secretion of key cell factors in HUCMSCs. These results demonstrate that the PET/PDA/dECM composite carrier holds great potential for scaling up MSCs' long-term in vitro cultivation and the production of paracrine factors.

Insights into FcγR involvement in pain-like behavior induced by an RA-derived anti-modified protein autoantibody

Joint pain is one of the most debilitating symptoms of rheumatoid arthritis (RA) and patients frequently rate improvements in pain management as their priority. RA is hallmarked by the presence of anti-modified protein autoantibodies (AMPA) against post-translationally modified citrullinated, carbamylated and acetylated proteins. It has been suggested that autoantibody-mediated processes represent distinct mechanisms contributing to pain in RA. In this study, we investigated the pronociceptive properties of monoclonal AMPA 1325:01B09 (B09 mAb) derived from the plasma cell of an RA patient. We found that B09 mAb induces pain-like behavior in mice that is not associated with any visual, histological or transcriptional signs of inflammation in the joints, and not alleviated by non-steroidal anti-inflammatory drugs (NSAIDs). Instead, we found that B09 mAb is retained in dorsal root ganglia (DRG) and alters the expression of several satellite glia cell (SGC), neuron and macrophage-related factors in DRGs. Using mice that lack activating FcγRs, we uncovered that FcγRs are critical for the development of B09-induced pain-like behavior, and partially drive the transcriptional changes in the DRGs. Finally, we observed that B09 mAb binds SGC in vitro and in combination with external stimuli like ATP enhances transcriptional changes and protein release of pronociceptive factors from SGCs. We propose that certain RA antibodies bind epitopes in the DRG, here on SGCs, form immune complexes and activate resident macrophages via FcγR cross-linking. Our work supports the growing notion that autoantibodies can alter nociceptor signaling via mechanisms that are at large independent of local inflammatory processes in the joint.

The predictive values of soluble B7-DC in children with refractory mycoplasma pneumoniae pneumonia.

Refractory mycoplasma pneumoniae pneumonia (RMPP) is a serious mycoplasma pneumoniae infection and is difficult to diagnose early. The levels of serum soluble B7-dendritic cell (sB7-DC) in children with mycoplasma pneumoniae pneumonia (MPP) were assessed to explore the clinical significance of sB7-DC levels in RMPP. A total of 65 patients with mycoplasma pneumoniae pneumonia (MPP) were enrolled in this study between January 2017 and December 2018. The patients were divided into the general mycoplasma pneumoniae pneumonia (GMPP) (n=30) and RMPP groups (n=35); the data of 20 normal children served as a control group (n=20). An enzyme-linked immunoassay kit was used to detect the expression of soluble B7-dendritic cell (sB7-DC) and other inflammatory factors. Binary logistic regression was performed to identify the independent predictors of RMPP. Receiver operating characteristic (ROC) curves were drawn to evaluate the value of each independent risk factor in the early diagnosis of RMPP. The results showed that compared to the GMPP group, children in the RMPP group had a significantly longer hospital stay and had a significantly longer fever duration (P<0.05). The values of interferon-gamma (IFN-γ), interleukin 17 (IL-17), and sB7-DC in the RMPP group were significantly higher than those in the normal control and GMPP groups (all P<0.05). The results of the correlation analysis showed that sB7-DC was positively correlated with IFN-γ and IL-17 and these indicators could be used in combination to evaluate the severity of the disease. The binary logistic regression analysis identified IL-17 and sB7-DC as independent risk factors for RMPP (P<0.05). The ROC curve analysis showed that the cut-off values of IL-17 and sB7-DC were 309.6 pg/L and 1,109.7 pg/mL, respectively. The areas under the curve (AUCs) of IL-17 and sB7-DC were 0.741 and 0.794, respectively. The sensitivity of IL-17 to RMPP prediction was 83.3%, and the specificity was 62.9%. The sensitivity and specificity of sB7-DC to RMPP were 86.7% and 62.9%, indicating that sB7-DC had the highest predictive power for RMPP. The level of serum sB7-DC may play an important role in the early diagnosis of RMPP. Our research results provide a theoretical basis for the early diagnosis of RMPP.

Human Cytomegalovirus and Human Herpesvirus 6 Coinfection of Dermal Fibroblasts Enhances the Pro-Inflammatory Pathway Predisposing to Fibrosis: The Possible Impact on Systemic Sclerosis.

Systemic sclerosis (SSc) is a severe autoimmune disease likely triggered by genetic and environmental factors, including viral infections. Human cytomegalovirus (HCMV) and human herpesvirus 6A species (HHV-6A) have been associated with SSc, based on in vivo and in vitro evidence, but the data are still inconclusive. Furthermore, despite both viruses being highly prevalent in humans and able to exacerbate each other’s effects, no data are available on their joint effects. Hence, we aimed to study their simultaneous impact on the expression of cell factors correlated with fibrosis and apoptosis in in vitro coinfected fibroblasts, representing the main target cell type in SSc. The results, obtained by a microarray detecting 84 fibrosis/apoptosis-associated factors, indicated that coinfected cells underwent higher and more sustained expression of fibrosis-associated parameters compared with single-infected cells. Thus, the data, for the first time, suggest that HCMV and HHV-6A may cooperate in inducing alterations potentially leading to cell fibrosis, thus further supporting their joint role in SSc. However, further work is required to definitively answer whether β-herpesviruses are causally linked to the disease and to enable the possible use of targeted antiviral treatments to improve clinical outcomes.

Effects of ursolic acid on oxidative stress and inflammatory factors in a rat model of AR after PM2.5 exposure

Objective: To investigate the effects of ursolic acid (UA) on oxidative stress and inflammatory factors in a rat model of AR after PM2.5 exposure. Methods: Sixty healthy female SD rats were randomly divided into five groups: normal control group (NC group), PM2.5 unexposed AR group (AR group), PM2.5 exposed AR group (ARE group), UA intervention AR group (AR+UA group), and UA intervention PM2.5 exposed AR group (ARE+UA group), with 12 rats in each group. AR model was performed by a basal sensitization with intraperitoneal injection of ovalbumin (OVA) and followed by nasal instillation. PM2.5 exposure was carried out by inhalation exposure system at a concentration of 200 μg/m3 for 3 h/d for 30 days. UA intervention group was given UA intragastric administration at 20 mg/(kg·d). AR symptoms including sneezing, nasal scratching and nasal secretion of rats in each group were observed. The activities of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in nasal mucosa were tested. The pathological changes of nasal mucosa were observed by HE staining. The levels of OVA-sIgE, IL-6 and IL-17 in serum were measured by enzyme-linked immunosorbent assay (ELISA). Protein microarray was used to measure the expression of multiple inflammation cell factors in nasal mucosa. Statistical analysis was performed with SPSS 20.0. Results: After UA intervention, the frequency of nasal sneezing, scratching and nasal secretion in ARE+UA group were lower than those of ARE group (P<0.05). Pathological examination of nasal mucosa showed that ARE+UA group had less inflammatory granulocyte infiltration and less pathological damage to the epithelial layer than ARE group. The activities of SOD in nasal mucosa of ARE+UA group were higher than those of ARE group ((50.10±3.09) U/mg vs (20.13±1.30) U/mg, F value was 597.54, P<0.01). The contents of MDA in nasal mucosa of ARE+UA group were lower than those of ARE group ((57.78±12.36) nmol/g vs (124.12±9.40) nmol/g, F value was 115.51, P<0.01). The expression levels of OVA-sIgE, IL-6 and IL-17 proteins were lower in the ARE+UA group than those in ARE group ((11.61±0.27) ng/ml vs (20.30±0.67) ng/ml, (47.59±15.49) pg/ml vs (98.83±10.98) pg/ml, (623.30±8.75) pg/ml vs (913.32±9.06) pg/ml, F value was 283.42, 80.45, 683.73, respectively, all P<0.01). After UA intervention, protein microarray analysis showed that the expression of IL-4, IL-6, IL-13, chemokine CXCL7, IL-1α, IL-1β, MMP-8 and MCP-1 in ARE+UA group was decreased compared with ARE group while IFN-γ and IL-10 increased (all P<0.01). Conclusion: UA can reduce the aggravated AR symptoms and pathological damage of nasal mucosa, inhibit oxidative stress and release of inflammatory factors after PM2.5 exposure, and thus plays a protective role in the pathological damage of AR induced by PM2.5 exposure.

A New Immortalized Human Alveolar Epithelial Cell Model to Study Lung Injury and Toxicity on a Breathing Lung-On-Chip System.

The evaluation of inhalation toxicity, drug safety and efficacy assessment, as well as the investigation of complex disease pathomechanisms, are increasingly relying on in vitro lung models. This is due to the progressive shift towards human-based systems for more predictive and translational research. While several cellular models are currently available for the upper airways, modelling the distal alveolar region poses several constraints that make the standardization of reliable alveolar in vitro models relatively difficult. In this work, we present a new and reproducible alveolar in vitro model, that combines a human derived immortalized alveolar epithelial cell line (AXiAEC) and organ-on-chip technology mimicking the lung alveolar biophysical environment (AXlung-on-chip). The latter mimics key features of the in vivo alveolar milieu: breathing-like 3D cyclic stretch (10% linear strain, 0.2 Hz frequency) and an ultrathin, porous and elastic membrane. AXiAECs cultured on-chip were characterized for their alveolar epithelial cell markers by gene and protein expression. Cell barrier properties were examined by TER (Transbarrier Electrical Resistance) measurement and tight junction formation. To establish a physiological model for the distal lung, AXiAECs were cultured for long-term at air-liquid interface (ALI) on-chip. To this end, different stages of alveolar damage including inflammation (via exposure to bacterial lipopolysaccharide) and the response to a profibrotic mediator (via exposure to Transforming growth factor β1) were analyzed. In addition, the expression of relevant host cell factors involved in SARS-CoV-2 infection was investigated to evaluate its potential application for COVID-19 studies. This study shows that AXiAECs cultured on the AXlung-on-chip exhibit an enhanced in vivo-like alveolar character which is reflected into: 1) Alveolar type 1 (AT1) and 2 (AT2) cell specific phenotypes, 2) tight barrier formation (with TER above 1,000 Ω cm2) and 3) reproducible long-term preservation of alveolar characteristics in nearly physiological conditions (co-culture, breathing, ALI). To the best of our knowledge, this is the first time that a primary derived alveolar epithelial cell line on-chip representing both AT1 and AT2 characteristics is reported. This distal lung model thereby represents a valuable in vitro tool to study inhalation toxicity, test safety and efficacy of drug compounds and characterization of xenobiotics.

Modulation of microRNome by Human Cytomegalovirus and Human Herpesvirus 6 Infection in Human Dermal Fibroblasts: Possible Significance in the Induction of Fibrosis in Systemic Sclerosis.

Human cytomegalovirus (HCMV) and Human herpesvirus 6 (HHV-6) have been reportedly suggested as triggers of the onset and/or progression of systemic sclerosis (SSc), a severe autoimmune disorder characterized by multi-organ fibrosis. The etiology and pathogenesis of SSc are still largely unknown but virological and immunological observations support a role for these beta-herpesviruses, and we recently observed a direct impact of HCMV and HHV-6 infection on the expression of cell factors associated with fibrosis at the cell level. Since miRNA expression has been found profoundly deregulated at the tissue level, here we aimed to investigate the impact on cell microRNome (miRNome) of HCMV and HHV-6 infection in in vitro infected primary human dermal fibroblasts, which represent one of the main SSc target cells. The analysis, performed by Taqman arrays detecting and quantifying 754 microRNAs (miRNAs), showed that both herpesviruses significantly modulated miRNA expression in infected cells, with evident early and late effects and deep modulation (>10 fold) of >40 miRNAs at each time post infection, including those previously recognized for their key function in fibrosis. The correlation between these in vitro results with in vivo observations is strongly suggestive of a role of HCMV and/or HHV-6 in the multistep pathogenesis of fibrosis in SSc and in the induction of fibrosis-signaling pathways finally leading to tissue fibrosis. The identification of specific miRNAs may open the way to their use as biomarkers for SSc diagnosis, assessment of disease progression and possible antifibrotic therapies.

Exercise interventions for post-stroke depression: A protocol for systematic review and meta-analysis.

Background:Post-stroke depression (PSD) is one of the most common neuropsychiatric complications after stroke and is associated with increased risk of death and poor functional outcomes. Strong evidence shows that exercise has benefits for depression. However, it is not clear whether exercise has benefits specifically for PSD. This study aims to explore the effects of exercise on PSD and to establish safe and effective exercise prescriptions.Methods and analysis:The PubMed, Cochrane Library and EMBASE, databases will be searched using prespecified search strategies. Randomized controlled trials and non-randomized prospective controlled cohort studies regarding exercise for PSD will be included. The primary outcomes are depression scale and stroke outcome. The secondary outcomes are the occurrence of adverse events, cognitive function, quality of life indices, and the expression of nerve cell factors. The methodological quality of each study will be evaluated by the physiotherapy evidence database scale. The heterogeneity will be evaluated using the I2 test. If I2 > 50%, random effects models will be used in the analysis; otherwise, fixed effects models will be used to pool the data.Results:This study will assess the efficacy and safety of exercise for PSD.Conclusions:Our findings will be helpful for clinicians to re-examine the clinical decision-making in the treatment of PSD, by assessing the efficacy of a promising treatment modality for patients with PSD.Ethics and dissemination:Ethical approval is not required because this study is a secondary analysis. The results of this study will be disseminated through journals and academic exchanges.Systematic review registration number:INPLASY202110100.

Wound-healing acceleration of mice skin by Sipunculus nudus extract and its mechanism

In order to explore the effect of Sipunculus nudus extract (SNE) on skin wound healing in mice and its mechanism, hemostasis effect of SNE was measured, the mouse skin wound model was established by full-thickness excision. The morphological changes of the wound were observed after the treatment with SNE and the healing rate was measured. The changes of wound histology were observed by hematoxylin eosin (HE) staining, Masson staining and transmission electron microscope (TEM). The expression of cell factors and related proteins was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results showed that the SNE possessed hemostatic function. SNE could obviously improve the healing rate of wound in mouse and shorten time of scab removal compared with the none-treatment (NT) group ( P < 0.05).The pathological histology analysis results showed complete epidermal regeneration, with remarkable capillary and collagen fiber observed in the SNE group. The expression level of tumor necrosis factor-α (TNF -α), interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) in SNE group was significantly lower than that of the NT group on 7 d ( P < 0.05). Moreover, compared with the NT group, the gene expressions level of Smad7 was significantly increased and the level of type II TGF-β receptors (TGF-βRII), collagen I (COL1A1) and α-smooth muscle actin (α-SMA) were significantly reduced in the SNE group on 28 d ( P < 0.05), but the difference was not statistically significant compared to Yunnanbaiyao group (PC group) ( P > 0.05). These results indicated that SNE possessed obvious activity of accelerating wound healing and inhibiting scar formation, and its mechanism was closely related to hemostatic function, regulation of inflammatory factors, collagen deposition, collagen fiber remodeling and intervening TGF-β/Smads signal pathway. Therefore, SNE may have promising clinical applications in skin wound repair and scar inhibition.

The effect of NK-1R-siRNA on expression of inflammatory factors in allergic rhinitis

Objective: To explore the effect of neurokinin-1 receptor small interfering RNA (NK-1R-siRNA) on the expression of inflammation factors in allergic rhinitis (AR). Methods: Twenty-four male SD rats were divided into three groups randomly (by random number table methord): NK-1R-siRNA group, negative control siRNA (NC-siRNA) group and saline group, with 8 rats in each group. SD rats were sensitized and challenged with ovalbumin (OVA) to induce AR. The rats were treated intranasally with NK-1R-siRNA, NC-siRNA or saline before and during the challenge period. The AR symptoms were observed. The levels of OVA-specific IgE were measured by enzyme-linked immunosorbent assay (ELISA). The levels of NK-1R expression in the nasal mucosal tissues were determined by real time PCR (RT-PCR) and immunohistochemistry. Antibody array was used in studying the expression of inflammation cell factors in nasal mucosa. SPSS 11.0 software was used for one-factor analysis of variance. Results: Compared with saline group, AR symptoms relived significantly in NK-1R-siRNA group (nose rubbing (31.4±8.9)/15 min vs (69.5±17.9)/15 min, sneezing (7.2±1.9)/15 min vs (23.7±9.2)/15 min, nasal secretions (7.1±2.3) mg vs (24.1±4.4) mg, t value was 38.100, 17.125, 16.837, respectively, all P<0.01), and the level of serum OVA-specific IgE was also reduced ((8.56±0.73) ng/ml vs (18.05±1.22) ng/ml, t=9.787, P<0.01). The RT-PCR and immunohistochemistry results showed that the expression of NK-1R in nasal mucosa of NK-1R-siRNA group was remarkably reduced than that of the NC-siRNA group and saline group. After the treatment of NK-1R-siRNA, the expression of interleukin (IL) 1α, IL-1β, IL-4, IL-6 and IL-13 decreased, while the interferon-γ (IFN-γ) and IL-10 increased. Conclusion: NK-1R-siRNA could regulate the release of inflammation factors in AR nasal mucosa, thus relive the allergic inflammation.

A Transforming Growth Factor-β and H19 Signaling Axis in Tumor-Initiating Hepatocytes That Regulates Hepatic Carcinogenesis.

Functions of transforming growth factor-β (TGF-β) in the liver vary depending on specific cell types and their temporal response to TGF-β during different stages of hepatocarcinogenesis (HCG). Through analysis of tumor tissues from hepatocellular carcinoma (HCC) patients, we were able to cluster hepatic epithelial cell-derived TGF-β gene signatures in association with distinct clinical prognoses. To delineate the role of hepatic epithelial TGF-β signaling in HCC development, we used an experimental system in which tumor-initiating hepatocytes (TICs) were isolated from TGF-β receptor II floxed mice (Tgfbr2fl/fl ) and transplanted into syngeneic C57BL/6J mice by splenic injection. Recipient mice were then administered Cre-expressing adenovirus (Ad-Cre) to inactivate Tgfbr2 in transplanted TICs. After latency, Tgfbr2-inactivated TICs formed larger and more tumor nodules in recipient livers compared to TICs without Tgfbr2 inactivation. In vitro analyses revealed that treatment of cultured TICs with TGF-β inhibited expression of progenitor cell factors (including SRY (sex determining region Y)-box 2 [Sox2]). RNA sequencing (RNA-seq) analysis identified H19 as one of the most up-regulated long noncoding RNA (lncRNA) in association with Tgfbr2 inactivation in TICs. Tgfbr2 inactivation by Ad-Cre led to a 5-fold increase of H19 expression in TICs. Accordingly, TGF-β treatment reduced H19 expression. We observed that forced overexpression of Sox2 in TICs increased transcription of H19, whereas knockdown of Sox2 decreased it. Furthermore, depletion of H19 reduced the progenitor property of TICs in vitro and decreased their tumorigenic potential in vivo. Finally, we observed a low level of H19 mRNA expression in human HCC tissues from patients with the epithelial TGF-β gene signature in association with favorable prognosis. Conclusion: Our findings describe a TGF-β and H19 signaling axis by Sox2 in TICs that importantly regulates HCG.

Effects of the modified electric convulsive treatment (MECT) on cell factors of schizophrenia.

The expression of cell factors of schizophrenia and the effect of the modified electric convulsive treatment (MECT) were studied. In total, 156 patients with schizophrenia were selected, and divided into the drug group (70 cases) and the drug combined with MECT group (combined group) (86 cases) according to the treatment methods. In addition, 70 cases of healthy volunteers (control group) were selected according to the closest matching method based on 1:1 of age and gender. The drug treatment, consisted of anti-psychotic drugs, such as risperidone 2–8, quetiapine 300–750, ziprasidone 80–160 or aripiprazole 10–30 mg/day, and for the control group, we used the electric spasm therapeutic instrument, Thymatron®IV Systems up to 6 times, 3 times a week. The levels of interleukin (IL)-10, IL-4, IL-6 and IL-1 were detected before and after treatment by ELISA. The positive and negative symptom scale (PANSS) was used to evaluate the efficiency. Before the treatment, IL-1 and IL-6 levels of drug and combined groups were significantly higher than those of the control group (P<0.05), while IL-4 and IL-10 had no difference with the control group. There was no significant difference of each factor between the drug and combined groups. After treatment, IL-1, IL-6 and IL-10 of the drug group did not change compared to the levels before treatment, but IL-4 increased significantly; IL-1 and IL-10 of the combined group did not change, while IL-4 and IL-6 increased significantly; IL-1, IL-4 and IL-6 of the drug and combined groups were significantly (P<0.05) higher than those in the control group, but not IL-10. IL-1, IL-4 and IL-6 levels of the combined group were significantly higher (P<0.05) than those of the drug group. After treatment, the PANSS scores of the two groups decreased and the combined group decreased more significantly (P<0.05). The reduction rate of the combined group was significantly higher (P<0.05) than that of the drug group. The total efficiency of the combined group was significantly higher than that of the drug group, and after comparing these levels, there was statistical significance (P<0.05). IL-1, IL-4, IL-6 and IL-10 levels of the drug and combined groups before treatment were not associated with PANSS scores and the variation of IL-1, IL-4, IL-6 and IL-10 of the drug and combined groups had no correlation with the reduction rate of the PANSS. The results showed that, cell factors of schizophrenia had an abnormal expression, and medication and MECT can affect the expression level. In addition, MECT can improve the effect in the treatment of schizophrenia, but had no obvious correlation with the change of cell factors.

Maintenance of a Schwann-Like Phenotype in Differentiated Adipose-Derived Stem Cells Requires the Synergistic Action of Multiple Growth Factors.

Differentiating human adipose-derived stem cells (ASCs) towards Schwann cells produces an unstable phenotype when stimulating factors are withdrawn. Here, we set out to examine the role of glial growth factor 2 (GGF-2) in the maintenance of Schwann-like cells. Following ASC differentiation to Schwann-like cells, stimulating factors were withdrawn such that cells either remained in media supplemented with all stimulating factors, GGF-2 alone, or underwent complete withdrawal of all factors. Furthermore, each stimulating factor was also removed from the growth medium individually. At 72 hours, gene (qRT-PCR) and protein (ELISA) expression of key Schwann cell factors were quantified and cell morphology was analysed. Cells treated with GGF-2 alone reverted to a stem cell morphology and did not stimulate the production of brain-derived neurotrophic factor (BDNF), regardless of the concentration of GGF-2 in the growth medium. However, GGF-2 alone increased the expression of Krox20, the main transcription factor involved in myelination, relative to those cells treated with all stimulating factors. Cells lacking fibroblast growth factor were unable to maintain a Schwann-like morphology, and those lacking forskolin exhibited a downregulation in BDNF production. Therefore, it is likely that the synergistic action of multiple growth factors is required to maintain Schwann-like phenotype in differentiated ASCs.

Modification of cell differentiation, one of the mechanisms in the surveillance of malignancy.

Most humans carry the potentially life-endangering Epstein-Barr virus (EBV). The immediate danger after infection is imposed by proliferation of the B cells that carry the viral genome. Although a number of different cell types can be infected with EBV, B lymphocytes are exceptionally sensitive; they express a set of virus-encoded proteins, which collaborate with host proteins to induce proliferation. This phenomenon can be demonstrated in vitro with experimentally infected B cells. These viral genes are expressed only in B lymphocytes and are restricted to a defined differentiation stage. This limitation is of high importance for the maintenance of the controlled EBV-carrier state of humans. The emergence of EBV-induced B-cell malignancies is counteracted by highly efficient immunologic mechanisms. Recognition of EBV-transformed immunoblasts in an MHC class I-restricted manner by cytotoxic CD8 T cells and, to a lesser extent, by CD4 T cells, is thought to play the major role. The in vitro experimental results are in accordance with the emergence of EBV(+) B-cell malignancies in immunosuppressive conditions. In this Masters primer, we emphasize that in addition to eliminating B cells that carry the virus genome, the regulatory circuit of the immune response also operates in surveillance, particularly in the early phase of infection. This mechanism involves T-cell-mediated regulation of B-cell differentiation. Because of the strict dependence of the viral growth program on the expression of host cell factors, altering the differentiation state can curb the proliferation of B cells that harbor the viral genome.

Changes in expression of hepatic fibrosis related cell factors in rats with bile duct obstruction

Changes in expression of hepatic fibrosis related cell factors in rats with bile duct obstruction

The effect of alendronate on the expression of important cell factors in osteoclasts

This study investigated the effects of alendronate (ALN) on critical cell factors in osteoclasts. RAW 264.7 cells were induced by sRANKL to change to mature osteoclasts. On the sixth day of incubation, the osteoclasts were treated with ALN at various concentrations and for different incubation times. The concentration groups included 10-5 M, 10-6 M and 10-7 M ALN, respectively. The cells were incubated for 0 (control group), 2, 4, 6 and 8 h for each dose group. The mRNA and protein expression of tartrate‑resistant acid phosphatase, carbonic anhydrase II, osteoclast‑associated receptor and FAS/FASL genes in osteoclasts was analyzed. A concentration- and time‑dependent decrease in the mRNA and protein expression levels of the five genes was observed, and no significant difference between the two control groups was observed (P>0.05). Notably, significant differences between any two experimental groups were observed (P<0.05). Thus, ALN significantly decreased the expression of critical factors involved in osteoclast function.

Influence of monochromatic light exposure on the growth of Miiller cell and the expression of myopia-related cell factors in vitro

Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Muller cells participate in the formation of myopia.However,the effect and mechanism of retinal Muller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Muller cells and the expression of cell factors in Muller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Muller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Muller cells were cultured with DMEM containing 10％ fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Muller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Muller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P＞0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Muller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia. Key words: Monochromatic light; Muller cell; Myopia; Transforming growth factor-beta 1 ; Tyrosine hydroxylase; Inducible nitric oxide synthase; Basic fibroblast growth factor

Influence of intravitreal avastin on the expression of cell factors in retinal proliferative membrane in proliferative diabetic retinopathy eye

Background A main cause of visual impairment in proliferative diabetic retinopathy (PDR) is vitreous hemorrhage and retinal detachment due to contraction of fibrovascular membrane.To explore the pathogenic mechanism of fibrovascular membrane is a new target for the prevention and management of PDR.Objective This study was to determine the change in expression of vascular endothelial growth factor (VEGF),connective tissue growth factor(CTGF) and pigment epithelium derived factor(PEDF) in the proliferative membranes of patients with PDR after intravitreal injection of avastin,an anti-VEGF agent.Methods This study was approved by the Medical Ethic Committee of Tianjin Medical College,and written informed consent was obtained from each patient before enrollment.A prospective randomized-controlled study was designed.Twenty-six eyes of 24 patients with PDR scheduled for surgery were enrolled from January to June,2008 in Tianjin Medical College Eye Hospital.The patients were randomized into the simple vitrectomy group and avastin injection combined with vitrectomy group,with matched gender,age and disease duration.1.25 mg (0.05 ml) of avastin was intravitreally injected prior to surgery,and vitrectomy was performed 10 days after injection in the avastin injection combined with vitrectomy group,and only vitrectomy was given in the simple vitrectomy group.Preretinal membrane was collected during the surgery.Expression of VEGF,CTGF and PEDF in the preretinal membranes was assayed by immunochemistry.Results VEGF,CTGF and PEDF were expressed in the cytoplasm.The rate of VEGF expression in the preretinal membranes was 30.77％ in the avastin injection combined with vitrectomy group,showing a significant reduction in comparison with the simple vitrectomy group(100.00％)(U =4.000,P＜0.01).The rate of expression CTGF was remarkable elevated in the avastin injection combined with vitrectomy group compared with the simple vitrectomy group (92.31％ vs.62.54％)(U=7.500,P=0.048).However,no significant difference was found in the expression rate of PEDF between the two groups(100.00％ vs.92.31％) (U =65.500,P =0.299).Conclusions The results suggest that intravitreal injection of anti-VEGF drugs resulted in the decrease of VEGF expression and increased CTGF expression in proliferative membranes from patients with PDR. Key words: Anti-vascular endothelial growth factor/avastin; Diabetic retinopathy; Retinal proliferative membrane; Vascular endothelial growth factor; Connective tissue growth factor; Pigment epithelium derived factor

Affection of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment

Objective To investigate the effect of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment,and then to discuss if it's effect on endometrial receptivity through its controlling the expression of cell factors which related to endometrial receptivity.Methods Mice's endometrium tissue samples and uterine Natural Killer (UNK) cells were obtained from pregnant mice on gd 4.5 days and gd 12.5 days.After endometrial glandular epithelial cells,endometrial stromal cells and UNK cells were separated through primary culture,epithelial cells and endometrial stromal cells at rate of 1 ∶ 1 a certain concentration UNK were mixed together and highly purified-cultured.Different concentrations of Zidan decoction was acted on the culture solution.FBS,black and aspirin control group were setup for contrast.The expression of LIF and VEGF which influenced by Zidan decoction in the immunity environment was assayed by Realtime-PCR.Results The expression of LIF and VEGF was significantly increased in the immunity environment of uterine (in 10％ Zidan decoction group,10％ aspirin group and 10％ blank control group:LIF was 2.10±0.20,1.98±0.14,0.90±0.05 separately,and VEGF was 3.17±0.31,2.52±0.09,0.92±0.06respectively,P＜0.05).Conclusion Zidan decoction could improve endometrial receptivity by increasing the expression of cell factor which related to endometrial receptivity with the help ofUNK paracrine. Key words: Zidan Decoction; Endometrial receptivity; Uterine Natural Killer cells; Co-culture

Identification of Ovarian Genes Regulated by Follicle-Stimulating Hormone During Early Secondary Oocyte Growth in Salmon.

It is well established in mammals that follicle-stimulating hormone (FSH) plays a major role in regulating oocyte growth and development by modulating the expression of follicle cell and oocyte factors. In fishes, however, little is known about the function(s) of FSH and regulation of early oocyte growth, a period of development that may affect later egg quality and fecundity. In species that exhibit synchronous gonad development (e.g., salmonids, catfish, striped bass), plasma FSH and/or pituitary gonadotropin subunit expression increases significantly during early secondary oocyte growth and continues to rise during vitellogenesis, suggesting FSH may play a role in regulating follicle development during this period. The goal of this study was to gain a fundamental understanding of how FSH acts at the level of the fish ovary by identifying FSH regulated ovarian genes. To examine time course effects of FSH, coho salmon (Oncorhynchus kisutch) previtellogenic ovarian follicles were cultured with or without 500 ng salmon FSH for 12-72 h. Culture media was collected for estradiol-17beta (E2) measurement, ovarian RNA was isolated, and abundance of candidate oocyte and follicle cell mRNAs was measured by quantitative PCR. Medium E2 levels were significantly elevated in response to FSH by 12 h and plateaued after 48 h. Messenger RNAs encoding steroidogenic enzymes, such as steroid acute regulatory protein (star) and 3beta-hydroxysteroid (hsd3b) were upregulated by 12 h, while P450 aromatase (cyp19a1a) showed no change in response to FSH. Other putative follicle cell transcripts such as FSH receptor (fshr) and lipoprotein lipase (lpl) were depressed initially by FSH treatment and later recovered to control levels. Putative oocyte transcripts such as zona pellucida glycoproteins, alveolin, and a C-type egg lectin showed no change over the 72-h experiment. Additional FSH regulated ovarian genes were subsequently identified through construction of four suppression subtractive hybridization (SSH) libraries with RNA isolated from FSH-treated and untreated whole ovarian follicles, as well as follicle/interstitial cell enriched ovarian tissues. Many of the genes identified by SSH are involved in signal transduction, cell cycle regulation, cell growth and proliferation, apoptosis, cholesterol metabolism and steroidogenesis. Experiments utilizing steroidogenesis inhibitors are underway to distinguish ovarian genes directly regulated by FSH versus those regulated by sex steroids produced in response to FSH. As evident by the rapid and significant increase in E2 release and changes in steroidogenic enzyme gene expression within the ovary, we can conclude that FSH strongly influences steroidogenesis of the salmon ovary during early secondary oocyte growth and may be a critical signal for puberty onset. This is the first comprehensive assessment of the biological actions of FSH in the fish ovary. Discoveries will aid in developing methods to reduce common reproductive dysfunctions in aquaculture, such as delayed puberty and poor egg quality. This project was supported in part by National Research Initiative Competitive Grant no. 2007-35203-18082 from the USDA Cooperative State Research, Education, and Extension Service to JAL. (platform)